BACKGROUND:The micro/nanostructured gradient biomimetic surface of implant materials can simulate the structure of the extracellular environment in human bone tissue,thereby achieving perfect bone integration function.However,further research is needed on the mechanisms by which the surface microstructure of bone implant materials regulates cell function and promotes osteogenesis. OBJECTIVE:To analyze the effect of titanium sheet microstructure surface on osteogenic differentiation of MC3T3-E1 osteoblasts. METHODS:(1)At a constant voltage of 5 V or 20 V,nanotube arrays of different diameters were prepared on the surface of titanium sheets by acid etching and anodic oxidation techniques,and were recorded as group R5 and group R20,respectively.The surface morphology,roughness,and hydrophilicity of pure titanium sheet(without acid etching or anodizing treatment)were measured in group R5 and group R20.(2)MC3T3-E1 osteoblasts of logarithmic growth stage were inoculated on the surface of pure titanium sheets,R5 group and R20 group respectively.After 24 hours of osteogenic induction culture,the expression of mechanical sensitive channel protein 1 was analyzed by RT-PCR and immunofluorescence staining.Osteoblast inducible base with or without the mechanosensitive channel protein 1 activator Yada1 was added,and alkaline phosphatase staining was performed after 7 days of culture.Alizarin red staining was performed after 14 days of culture. RESULTS AND CONCLUSION:(1)The surface of pure titanium sheets was smooth under scanning electron microscope.Relatively uniform and orderly nanotube arrays with average diameters of about 30 nm and 100 nm were observed on the surface of titanium sheets of groups R5 and R20,respectively.The results of scanning electron microscope were further verified by atomic force microscopy.The surface roughness of titanium sheet of group R5 was higher than that of pure titanium(P<0.05),and the water contact angle was lower than that of pure titanium(P<0.05).The surface roughness of titanium sheet in group R20 was higher than that in group R5(P<0.05),and the water contact angle was lower than that in group R5(P<0.05).(2)RT-PCR and immunofluorescence staining showed that the expression of mechanosensitive channel protein 1 in group R5 was higher than that in pure titanium group(P<0.05),and the expression of mechanosensitive channel protein 1 in group R20 was higher than that in group R5(P<0.05).Under the osteogenic induction,compared with the condition without Yada1,there were no significant changes in the activity of alkaline phosphatase and the deposition of calcified nodules in pure titanium group after Yada1 addition,while the activity of alkaline phosphatase and the deposition of calcified nodules in groups R5 and R20 after Yada1 addition were significantly increased(P<0.05).With or without Yada1,the alkaline phosphatase activity and calcified nodule deposition in group R5 were higher than those in pure titanium group(P<0.05),and the alkaline phosphatase activity and calcified nodule deposition in group R20 were higher than those in group R5(P<0.05).(3)The results show that the surface microstructure of titanium sheet can promote the osteogenic differentiation of osteoblast MC3T3-E1 by activating mechanosensitive channel protein 1.

Objective To explore the occurrence risk of enteral nutrition intolerance and analyze its influencing factors in 302 elderly critically ill patients. Methods The clinical case data of elderly critically ill patients in department of elderly cadres of the hospital were retrospectively analyzed from January 2019 to January 2024. According to the occurrence of enteral nutrition intolerance or not, they were divided into occurrence group (n=156) and non-occurrence group (n=146). The risk of nutritional intolerance in elderly critically ill patients was evaluated by feeding intolerance risk assessment form, and the influencing factors of enteral nutrition intolerance were analyzed by multivariate logistic regression analysis. Results Among the 302 elderly patients with critical illness, 53.31% (161/302) had high risk of enteral nutrition intolerance, and 51.66% (156/302) had enteral nutrition intolerance. Multivariate logistic analysis revealed that CRP level>10mg/L, APACHE-II score≥20 points, Lac≥3mmol/L and hypoalbuminemia were risk factors in elderly critically ill patients (OR=1.806, 2.977, 8.232, 3.031, P=0.011, 0.001, 0.041, 0.047), and addition of dietary fiber was a protective factor for enteral nutrition intolerance (OR=1.652, P=0.037). Conclusion The risk of enteral nutrition intolerance is high in elderly critically ill patients. Lac level, CRP level, hypoalbuminemia, and APACHE-II score of patients are independent risk factors for enteral nutrition intolerance, and addition of dietary fiber is a protective factor. It is necessary to take targeted interventions for patients according to the above factors to minimize the occurrence of enteral nutrition intolerance.

ObjectiveTo investigate the efficacy and mechanism of Polygoni Cuspidati Rhizoma et Radix （Huzhang） in treating respiratory syncytial virus（RSV） infection by regulating pulmonary surfactant lipid homeostasis through lipidomics. MethodsSixty BALB/c mice were randomly divided into the blank group， model group， positive group（ribavirin group， 46 mg·kg^-1）， and low- and high-dose Huzhang groups（0.75， 2.25 g·kg^-1）， with 12 mice in each group. Except for the blank group， all other groups were infected with RSV via intranasal instillation. The drug intervention groups were given corresponding doses of drug by gavage for 3 consecutive days， while normal saline was used in the blank and model groups. Hematoxylin-eosin（HE） staining was used to observe pathological changes in mouse lung tissue. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was employed to detect viral loads［RSV-nucleoprotein（N） and RSV-glycoprotein（G） mRNA］ and inflammatory factor levels［interleukin（IL）-1β and tumor necrosis factor（TNF）-α mRNA］ in the lung tissue. Mouse bronchoalveolar lavage fluid was collected to detect the levels of pulmonary surfactant lipids through ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive Orbitrap-MS）， followed by principal component analysis and differential lipid identification. ResultsCompared with the blank group， the model group exhibited extensive inflammatory cell infiltration， congestion， and tissue damage in the lungs， and the pathological score and lung index of lung tissue significantly increased（P<0.01）， along with significantly elevated mRNA expressions of RSV-N， RSV-G， IL-1β， and TNF-α（P<0.01）. Compared with the model group， different doses of Huzhang and ribavirin significantly reduced the pathological scores of the lung tissue and lung index（P<0.01）. In addition， the mRNA levels of RSV-N， RSV-G and TNF-α in the lungs significantly decreased in the Huzhang high dose group（P<0.01）. Lipidomics analysis identified multiple significantly changed differential metabolites. Compared with the blank group， the model group showed obvious abnormal lipid metabolism， which was manifested by the elevated levels of prostaglandin（PG）， ceramide（Cer）， phosphatidylcholine（PC）， phosphatidylethanolamine（PE）， phosphatidylinositol（PI）， sphingomyelin（SM）， and the decreased levels of diglycerides（DG） and acylethanolamine（NAE）. After the intervention of low dose of Huzhang， the above lipid metabolites showed a significant reversal trend， while the intervention of high dose of Huzhang could regulate levels of PI lipids， PG lipids and PC lipids. ConclusionHuzhang can significantly reduce the viral load of lung tissue and improve lung inflammation in RSV-infected mice. The underlying mechanism may be related to the maintenance of homeostasis in pulmonary surfactant lipids such as PI and PG.

ObjectiveTo investigate the efficacy and mechanism of Polygoni Cuspidati Rhizoma et Radix （Huzhang） in treating respiratory syncytial virus（RSV） infection by regulating pulmonary surfactant lipid homeostasis through lipidomics. MethodsSixty BALB/c mice were randomly divided into the blank group， model group， positive group（ribavirin group， 46 mg·kg^-1）， and low- and high-dose Huzhang groups（0.75， 2.25 g·kg^-1）， with 12 mice in each group. Except for the blank group， all other groups were infected with RSV via intranasal instillation. The drug intervention groups were given corresponding doses of drug by gavage for 3 consecutive days， while normal saline was used in the blank and model groups. Hematoxylin-eosin（HE） staining was used to observe pathological changes in mouse lung tissue. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was employed to detect viral loads［RSV-nucleoprotein（N） and RSV-glycoprotein（G） mRNA］ and inflammatory factor levels［interleukin（IL）-1β and tumor necrosis factor（TNF）-α mRNA］ in the lung tissue. Mouse bronchoalveolar lavage fluid was collected to detect the levels of pulmonary surfactant lipids through ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive Orbitrap-MS）， followed by principal component analysis and differential lipid identification. ResultsCompared with the blank group， the model group exhibited extensive inflammatory cell infiltration， congestion， and tissue damage in the lungs， and the pathological score and lung index of lung tissue significantly increased（P<0.01）， along with significantly elevated mRNA expressions of RSV-N， RSV-G， IL-1β， and TNF-α（P<0.01）. Compared with the model group， different doses of Huzhang and ribavirin significantly reduced the pathological scores of the lung tissue and lung index（P<0.01）. In addition， the mRNA levels of RSV-N， RSV-G and TNF-α in the lungs significantly decreased in the Huzhang high dose group（P<0.01）. Lipidomics analysis identified multiple significantly changed differential metabolites. Compared with the blank group， the model group showed obvious abnormal lipid metabolism， which was manifested by the elevated levels of prostaglandin（PG）， ceramide（Cer）， phosphatidylcholine（PC）， phosphatidylethanolamine（PE）， phosphatidylinositol（PI）， sphingomyelin（SM）， and the decreased levels of diglycerides（DG） and acylethanolamine（NAE）. After the intervention of low dose of Huzhang， the above lipid metabolites showed a significant reversal trend， while the intervention of high dose of Huzhang could regulate levels of PI lipids， PG lipids and PC lipids. ConclusionHuzhang can significantly reduce the viral load of lung tissue and improve lung inflammation in RSV-infected mice. The underlying mechanism may be related to the maintenance of homeostasis in pulmonary surfactant lipids such as PI and PG.

Objective To explore risk factors for cardiac valve calcification (CVC) in maintenance hemodialysis (MHD) patients and evaluate its impact on cardiovascular events and mortality. Methods Retrospective selection of 223 patients with MHD admitted to the Department of Nephrology of Jinshan Hospital, Fudan University from June 30, 2019 to June 30, 2024, and enrollment completed within one week of June 30, 2019. Patients were divided into CVC and non-CVC groups. Baseline data and 5-year follow-up data were collected. The binary logistic regression analysis was performed to explore the risk factors for CVC. Kaplan-Meier survival curve was used to analyze the survival rate of patients. Cox proportional hazard regression model was applied to evaluate the impact of CVC on the survival rates of MHD patients. Results Totally, 223 MHD patients with an average age of (58.4±13.5) years and an average dialysis duration of (64.0±55.4) months were involved. Among them, 136(61.0%) were males, 117(52.5%) were complicated with CVC. Age, dialysis duration, diabetic kidney disease (DKD), the serum corrected total calcium and phosphate, intact parathyroid hormone (iPTH), high-sensitive C-reactive protein (hsCRP), and homocysteine (Hcy) were independent related factors for CVC (P＜0.05). Both all-cause mortality (46.6% vs 28.7%) and cardiovascular mortality (33.3% vs 16.0%) were significantly higher in the CVC group than those in the non-CVC group (P＜0.01). Conclusions Age, dialysis duration, the primary disease, calcium and phosphate, and inflammation- and nutrition-related serum indicators are associated with CVC in MHD patients. CVC significantly increases mortality risk of MHD patients.

BACKGROUND: The purpose of this study was to evaluate the safety and efficacy of subsequent radiotherapy (RT) following first-line treatment with durvalumab plus chemotherapy in patients with extensive-stage small cell lung cancer (ES-SCLC). METHODS: A total of 122 patients with ES-SCLC from three hospitals during July 2019 to December 2021 were retrospectively analyzed. Inverse probability of treatment weighting (IPTW) analysis was performed to address potential confounding factors. The primary focus of our evaluation was to assess the impact of RT on progression-free survival (PFS) and overall survival (OS). RESULTS: After IPTW analysis, 49 patients received durvalumab plus platinum-etoposide (EP) chemotherapy followed by RT (Durva + EP + RT) and 72 patients received immunochemotherapy (Durva + EP). The median OS was 17.2 months vs . 12.3 months (hazard ratio [HR]: 0.38, 95% confidence interval [CI]: 0.17-0.85, P = 0.020), and the median PFS was 8.9 months vs . 5.9 months (HR: 0.56, 95% CI: 0.32-0.97, P = 0.030) in Durva + EP + RT and Durva + EP groups, respectively. Thoracic radiation therapy (TRT) resulted in longer OS (17.2 months vs . 14.7 months) and PFS (9.1 months vs . 7.2 months) compared to RT directed to other metastatic sites. Among patients with oligo-metastasis, RT also showed significant benefits, with a median OS of 17.4 months vs . 13.7 months and median PFS of 9.8 months vs . 5.9 months compared to no RT. Continuous durvalumab treatment beyond progression (TBP) prolonged OS compared to patients without TBP, in both the Durva + EP + RT (NA vs . 15.8 months, HR: 0.48, 95% CI: 0.14-1.63, P = 0.238) and Durva + EP groups (12.3 months vs . 4.3 months, HR: 0.29, 95% CI: 0.10-0.81, P = 0.018). Grade 3 or 4 adverse events occurred in 13 (26.5%) and 13 (18.1%) patients, respectively, in the two groups; pneumonitis was mostly low-grade. CONCLUSION Addition of RT after first-line immunochemotherapy significantly improved survival outcomes with manageable toxicity in ES-SCLC.
Humans ; Small Cell Lung Carcinoma/therapy* ; Retrospective Studies ; Male ; Female ; Middle Aged ; Lung Neoplasms/therapy* ; Aged ; Antibodies, Monoclonal/therapeutic use* ; Adult ; Immunotherapy/methods* ; Aged, 80 and over

Intervention in chronically activated microglia-mediated neuroinflammation is a novel approach to treat Alzheimer's disease (AD). The low permeability of the blood‒brain barrier (BBB) and non-selective distribution in the brain severely restrict AD drugs' disease-modifying efficacy. Here, an immunosuppressant TREM2-lowing antisense oligonucleotides (ASOs) and resveratrol co-loaded cationic liposome is developed as an immune reprogramming nanomodulator modified by acid-cleavable BBB-targeting peptide and microglia-targeting peptide (Res@TcMNP/ASO) for AD management. Res@TcMNP/ASO can enter brain endothelial cells via D-T7 peptides. Then D-T7 undergoes an acid-responsive cleavage, facilitating the escape of Res@MNP/ASO from endo/lysosomes to cross the BBB. The detached Res@MNP/ASO specifically targets M1-phenotype microglia via exposed MG1 peptides to prompt the simultaneous delivery of two drugs into activated microglia. This nanomodulator can not only restore the immune function of microglia through TREM2-lowing ASO but also mitigate the immune stimulation to microglia caused by reactive oxygen species (ROS) through resveratrol, thereby synergistically inhibiting the chronic activation of microglia to alleviate neuroinflammation in AD. Our results indicate that this combination treatment can achieve significant behavioral and cognitive improvements in late APP/PS1 mice.

Objective To explore the comparative application of phase and diaphragmatic navigation in three-dimensional magnetic resonance cholangiopancreatography(3D-MRCP)thin-layer scanning in elderly patients.Methods A total of 180 elderly patients were scanned by phase and diaphragmatic navigation via Siemens Aera1.5T superconducting MR scanner.The acquired images were reconstructed by 3D reconstruction.The anatomical structure,image quality and disease diagnosis were compared between the phase and diaphragmatic navigation groups.Results In liver of anatomy,the liver of primary bile duct,the superior,middle and inferior extrahepatic bile duct and the gallbladder could be well displayed,and the difference was not statistically significant between the two groups(P＞0.05).The display of pancreatic duct and the liver of secondary bile duct of diaphragmatic navigation was significantly better than those of phase navigation(P＜0.05).In terms of image quality,the excellent rate of diaphragmatic navigation was significantly higher than that of phase navigation,and the difference was statistically significant(P＜0.05).There were no statistically significant differences in the detection rate of pancreatobiliary system diseases,the diagnostic rate of cholelithiasis,common bile duct stones,common bile duct dilatation and pancreatic duct dilatation between the two groups(P＞0.05).Conclusion Diaphragmatic navigation is signifi-cantly better than phase navigation in the display of the anatomical structure of the pancreatic duct,the liver of secondary bile duct,and the excellent rate of image quality.Diaphragmatic navigation is more suitable for thin-layer 3D-MRCP scanning in elderly patients.

Objective: To establish an optimized method for isolation and purification of microglia from aged rat brain tissue, and the phenotype of microglia was detected by flow cytometry. Methods: With young rats(3 months old) as control, the brain tissues of aged rats were immediately processed into single cell suspensions by mechanical dissociation and enzymatic digestion using type IV collagenase. Microglia were isolated on Percoll gradients(30%-37%-70%). The cells were stained with fluorescence-labeled antibodies and the phenotype of microglia was detected by flow cytometry. Results: This study developed a method that enzymatic digestion and mechanical dissociation combined with density gradient centrifugation. More single cells could be obtained by using this method. And the survival rate of cells was more than 90%. The flow cytometric analysis showed that the expression of M1 microglia marker CD86 and MHC Ⅱ increased(P<0.01), and the expression of M2 microglia marker CD200R increased(P<0.01) in aged rats compared with that in young rats. Conclusion The use of type IV collagenase and mechanical digestion combined with density gradient centrifugation is good for isolating and purifying microglia from adult and aged rat brain tissue.